Harvesting Arabidopsis Seeds
Ben Carter, 9/24/2014 Seed Hygiene Arabidopsis seeds will bounce around and also occasionally adhere to clothing and skin. It is imperative that you wipe down the work area as well as your hands and arms between collecting vials of seeds. Do not use lab mats on the work area as they adhere to seeds.Transgenic plant material must be discarded into biohazard bags, not the regular trash. The containers of 1dram seed collection vials are to remain sealed and covered during the collection process to prevent seeds from bouncing into them or their container. Seed Harvesting # Fold a sheet of printer paper into quarters such that the creases form a pool in the center of the paper. (In other words, both folds "up.") # Enter the information for the first pot of plants to be harvested into the database. In our lab, this is FileMaker Pro. # Examine whether the dried plants are staked and tied with one stake for bulking or on a plant-by-plant basis. Based on this, refer to the appropriate section below. Collecting Seeds in Bulk # Remove the stake from the dried plants by carefully twisting and lifting it. # Use scissors to cut the inflorescences approximately 1" above the level of the soil. # Hold the dried plants above the folded printer paper and fold them over in half. Gently roll the dried plant material in your hands, allowing the seeds to fall and collect on the paper. # Fold the plant material in half again and repeat Step 3. # Discard the plant material. Collect the dispersed seeds into the center of the folded paper. # Pinch off the dried plant material on top of the seeds and discard it. Gently shake the paper to allow more plant material to surface. # Hold the folded paper in front and above the biohazard trash. Carefully blow on the seeds to remove dried plant material. Gently shake the paper to re-orient the seeds and repeat this step until the seeds are relatively free of dried plant material. # Obtain a clean 1dram glass screw-cap vial. Place a crease of the folded printer paper against the top of the vial and allow the seeds to flow into the vial. If this is difficult, transfer the seeds to a smaller second piece of folded paper and try again. # Place the cap on the vial and label it with the new seed stock number you entered into the database. Print out a label for the side of the vial with the relevant information. Place the vial in the seed stock library. Clean your area using the seed hygiene guidelines above. Collecting Seeds from Individual Plants # Remove the stake from the first dried plant by carefully twisting and lifting it. # Carefully disentangle the first plant from the others by collecting separating its inflorescences from the neighboring plants at its base and then moving up the plant in the same manner. # Inspect the plant to ensure that no material from neighboring plants has separated and become entangled. This would lead to seed stock cross-contamination. # Follow the steps in the "Collecting Seeds in Bulk" section above for your individual plant starting from Step 2. Category:Protocols __NOEDITSECTION__ Category:Arabidopsis